Sodium 3-[[4-amino-9,10-dihydro-3-[2-(2-methoxyethoxy)ethoxy]-9,10-dioxo-1-anthryl]amino]-2,4,6-trimethylbenzenesulphonate

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2002-2003

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2003

Report date:

2003

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 429 (Skin Sensitisation: Local Lymph Node Assay)

Version / remarks:

adopted 24 June 2002

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Type of study:

mouse local lymph node assay (LLNA)

Test material

Test material information

Constituent 1

Reference substance name:

Sodium 3-[[4-amino-9,10-dihydro-3-[2-(2-methoxyethoxy)ethoxy]-9,10-dioxo-1-anthryl]amino]-2,4,6-trimethylbenzenesulphonate

EC Number:

274-366-7

EC Name:

Sodium 3-[[4-amino-9,10-dihydro-3-[2-(2-methoxyethoxy)ethoxy]-9,10-dioxo-1-anthryl]amino]-2,4,6-trimethylbenzenesulphonate

Cas Number:

70179-77-0

Molecular formula:

C28H30N2O8S.Na

IUPAC Name:

sodium 3-[[4-amino-9,10-dihydro-3-[2-(2-methoxyethoxy)ethoxy]-9,10-dioxo-1-anthryl]amino]-2,4,6-trimethylbenzenesulphonate

Test material form:

solid: particulate/powder

In vivo test system

Study design: in vivo (LLNA)

Vehicle:

other: ethanol:water, 7:3 (v/v)

Concentration:

5 %, 1 0 % and 25 %

No. of animals per dose:

4

Positive control substance(s):

hexyl cinnamic aldehyde (CAS No 101-86-0)

Results and discussion

Positive control results:

ALPHA-HEXYLCINNAMALDEHYDE showed an allergenic potency when tested at concentration of 25 % (w/v).
EC3 is the estimated concentration for a STIMULATION INDEX of 3. In this study EC3 of 11.3% (w/v) was theoretically calculated with STIMULATION INDICES of 2.6 and 7.1 at test item concentrations of 10 % and 25 % (w/v).

In vivo (LLNA)

Results

Key result

Parameter:

EC3

Value:

ca. 0.9

Variability:

Values of 0.9 at 5 and 25 % and of 0.8 at 10 %

Test group / Remarks:

all test groups

Any other information on results incl. tables

Test item concnetration		Measurement		Calculation		Result
% (w/v)		dpm	dpm - BG	number of nodes	dpm per node	S.l.
	BG I	1	-	-	- -	-
	BG II	1	-	-	-	-
	CG1	4100	4099	8	512
5	TG 2	3665	3664	8	458	0.9
10	TG3	3424	3423	8	428	0.8
25	TG4	3766	3765	8	471	0.9

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

In this LLNA study STIMULATION INDICES of 0.9, 0.8 and 0.9 were determined with the test item at concentrations of 5 %, 10 % and 25 % (w/v) in ethanol:water, 7:3 (v/v).
A test item is regarded as a sensitizer in the LLNA if the exposure to at least one concentration of the test item resulted in an incorporation of 3HTdR at least 3-fold or greater than that recorded in control mice, as indicated by the STIMULATION INDEX.
Based on these criteria, the test item was found to be a non-sensitizer when tested up to 25 % (w/v) in ethanol:water, 7:3 (v/v).

Executive summary:

In order to study a possible allergenic potential of Sanolin Violett E2R three groups of four female mice were each treated with the test item at concentrations of 5 %, 1 0 % and 25 % (w/v) in ethanol:water, 7:3 (v/v) by topical application to the dorsum of each ear lobe (left and right) on three consecutive days. A control group of four mice was treated with the vehicle (ethanol:water, 7:3 (v/v)) only. Five days after the first topical application the mice were injected intravenously into a tail vein with radio-labelled thymidine eH-methyl thymidine).

Approximately five hours after intravenous injection, the mice were sacrificed, the draining auricular lymph nodes excised and pooled per group. Single cell suspensions of lymph node cells were prepared from pooled lymph nodes which were washed subsequently and incubated with trichloroacetic acid overnight. The proliferative capacity of pooled lymph node cells was determined by the incorporation of 3H-methyl thymidine measured in a ß-scintillation counter.

No test item-related clinical signs were observed.

All treated animals survived the scheduled study period.

A test item is regarded as a sensitizer in the LLNA if the exposure to at least one concentration of the test item resulted in an incorporation of 3HTdR at least 3-fold or greater than that recorded in control mice, as indicated by the STIMULATION INDEX (S.I.).

In this study STIMULATION INDICES of 0.9, 0.8 and 0.9 were determined with the test item at concentrations of 5 %, 10 % and 25 % (w/v) in ethanol:water, 7:3 (v/v).

Based on these criteria, the test item Sanolin Violett E2R was found to be a non-sensitizer when tested up to 25 % (w/v) in ethanol:water, 7:3 (v/v).